Experimental Study Of The Effect Of Moxibustion "Zusanli" On Notch Signaling Pathway In Bone Marrow Cells Of Mice After Chemotherapy

Objective:Focusing on the theme of moxibustion to improve bone marrow suppression after chemotherapy and increase the level of peripheral blood leukocyte in healthy mice,this paper explores the changes of Notch signal transduction pathway after moxibustion treatment.To clarify whether the whitening effect of moxibustion is achieved by regulating this signal pathway system,and to detect the protein expression of important signal molecules in the Notch signaling pathway.The mechanism of moxibustion in treating myelosuppression after chemotherapy was discussed from the aspect of molecular biology in order to provide a scientific basis for the clinical application of moxibustion in the treatment of myelosuppression after tumor chemotherapy.Methods:Sixty-one SPF male Kunming mice were selected,with a mass of 18 to 20 g,6to 8 weeks of age and 5 days in the laboratory.The tumor mass was peeled off under the aseptic conditions of A549 lung cancer seed mice,and then the homogenate was prepared,and the number of cells was adjusted to 1-10⁷/ml,and 0.2 ml was inoculated under the armpit of 50 mice.Thirty-one tumor-bearing mice with similar tumor size were randomly divided into blank group,model group and moxibustion treatment group.The normal group of simulated inoculation was set up.After the grouping,the experimental mice were stained and numbered with picric acid.After that,the base white blood cell count was checked and the two-pole mice were deleted.Each group was reserved for 8 A mouse used in the experiment.In aseptic condition of Lewis lung cancer,the tumor pieces were removed and homogenized,and the number of cells was adjusted to 1×10 ^7/ml.0.2ml was inoculated under the axils of each mouse.Selection of tumor-bearing mice with similar tumor size on the7th day of inoculation,A myelosuppression mouse model was established by cyclophosphamide（CTX）according to the"compilation of guiding principles for preclinical Research of New drugs（Western Medicine）"（compiled by the Pharmaceutical Administration of the Ministry of Health of the people’s Republic of China）.The rats were injected intraperitoneally according to the dosage of 100mg/kg/d for 3 days.According to the pharmacokinetic process of cyclophosphamide,the drug activity disappeared 4 hours later（Zhongshan Medical College,et al.）.Basic pharmacology.People’s Health Publishing House,1979/442),the model was established.Mice in blank control group were injected with saline intraperitoneally according to the dosage of 0.02ml/g.In the moxibustion group,the experimental mice were fixed on the fixed plate of self-made mice,and the"Zusanli"point was taken for suspended moxibustion for 3 minutes each time,once a day for 5 days.Normal group,blank group,model group,daily binding and grasping stimulation,the course of treatment was the same as moxibustion group;after the end of the treatment,the mice were killed under anesthesia,eyeball blood was collected,and bilateral femurs were taken to be preserved for use.The experimental process strictly complies with the relevant regulations issued by the State Ministry of Science and Technology on guiding opinions on the good treatment of Laboratory Animals.After treatment,the effect of moxibustion on leukocytes was observed,the pathological changes of bone marrow tissue were observed by HE staining,the protein expression of related genes was detected by immunohistochemistry,and the expression level of Notch signal pathway related genes in chemotherapy mice was observed by moxibustion.The mechanism of moxibustion improving bone marrow suppression and leukocyte after chemotherapy was discussed from the aspect of signal transduction pathway of bone marrow cells.Results:1.The results of blood routine examination showed that there was no significant change in the number of white blood cells before and after treatment in the normal group,blank group and model group,in which the mice in the normal group and the blank group remained within the normal range.The mice in the model group were kept in a low level.The number of leukocytes in the moxibustion group was very small before treatment.With the increase of treatment time,the number of leukocytes gradually increased significantly and reached the normal level on the fifth day（P<0.05）.2.The results of HE pathological examination showed that the bone marrow tissue structure of the normal group and the blank group remained intact and the bone marrow cells were uniformly distributed in the model group,the bone marrow tissue structure of the model group was destroyed,the distribution of the bone marrow cells was disordered,and the cell density was decreased.In the treatment group,the bone marrow tissue structure returned to normal,the bone marrow cell density increased,and the structure was intact.3.Detection of CSL,Jagged1,Notch1 protein expression in bone marrow tissue of mice in each group showed that there was no difference between the normal group and the blank group in the change of CSL,Jagged1,Jagged2 protein in bone marrow tissue.Compared with the blank group,the expression of CSL,Jagged1,Jagged2 protein in the bone marrow tissue of the model group was significantly lower than that of the control group（P<0 05）.Compared with the model group,the expression of CSL,Jagged1,Jagged2 protein in bone marrow of the moxibustion group was significantly higher than that of the model group（P<0 05）.conclusion:1.Moxibustion can effectively improve white blood cell level after chemotherapy in mice2.The therapeutic mechanism of myelosuppression in mice after chemotherapy may be to activate the Notch signaling pathway by regulating some factors in the Notch signaling pathway,to enhance the hematopoietic function of bone marrow,and thus to regulate the number of hematopoietic cells in bone marrow.So as to achieve therapeutic effect.